Variants of interleukin-1 receptor antagonist: compositions and uses thereof

Abstract

The present invention provides Interleukin-1 receptor antagonist splice variants, including isolated nucleic acids encoding these variants and the encoded amino acid sequences, as well as antibodies, antisense oligonucleotides, expression vectors and host cells that include these sequences. The present invention further discloses methods of using these sequences in the diagnosis, prognosis, treatment, and prevention of diseases and disorders mediated by Interleukin-1.

1 . An isolated nucleic acid comprising a polynucleotide sequence encoding a polypeptide capable of binding to a mammalian IL-1 receptor which is at least 80% homologous to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and fragments, variants and analogs thereof.  
     
     
         2 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide which is at least 85% homologous to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and fragments, variants and analogs thereof.  
     
     
         3 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide which is at least 90% homologous to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and fragments, variants and analogs thereof.  
     
     
         4 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide which is at least 95% homologous to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and fragments, variants and analogs thereof.  
     
     
         5 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide which is 100% homologous to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and fragments, variants and analogs thereof.  
     
     
         6 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide comprising contiguous amino acids having at least 80% homology to positions 44 to 73 set forth in SEQ D NO:4.  
     
     
         7 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide comprising contiguous amino acids having at least 90% homology to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         8 . The isolated nucleic acid according to  claim 1  wherein the polynucleotide sequence encodes a polypeptide comprising contiguous amino acids having at least 95% homology to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         9 . The isolated nucleic acid according to  claim 1 , wherein the polynucleotide sequence encodes a polypeptide comprising contiguous amino acids having 100% identity to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         10 . An isolated nucleic acid comprising a polynucleotide sequence complementary to the polynucleotide sequence according to  claim 1 .  
     
     
         11 . An isolated nucleic acid comprising a polynucleotide sequence that hybridizes under stringent conditions to a polynucleotide sequence according to  claim 1 .  
     
     
         12 . An isolated nucleic acid comprising a polynucleotide sequence having a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 and SEQ ID NO:6.  
     
     
         13 . An isolated nucleic acid comprising a polynucleotide sequence complementary to the polynucleotide sequence according to  claim 12 .  
     
     
         14 . The polynucleotide of  claim 1  wherein the polypeptide encoded is devoid of at least one IL-1 activity.  
     
     
         15 . The polynucleotide of  claim 1  wherein the polypeptide encoded is substantially devoid of IL-1 activity.  
     
     
         16 . A polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 and fragments, variants and analogs thereof, wherein said polypeptide binds to a mammalian IL-1 receptor.  
     
     
         17 . A polypeptide which is at least 80% homologous to the polypeptide of  claim 16 .  
     
     
         18 . A polypeptide which is at least 85% homologous to the polypeptide of  claim 16 .  
     
     
         19 . A polypeptide which is at least 90% homologous to the polypeptide of  claim 16 .  
     
     
         20 . A polypeptide which is at least 95% homologous to the polypeptide of  claim 16 .  
     
     
         21 . The polypeptide according to  claim 16  having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.  
     
     
         22 . The polypeptide according to  claim 16  comprising contiguous amino acids having at least 80% homology to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         23 . The polypeptide according to  claim 16  comprising contiguous amino acids having at least 90% homology to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         24 . The polypeptide according to  claim 16  comprising contiguous amino acids having at least 95% homology to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         25 . The polypeptide according to  claim 16  comprising contiguous amino acids having 100% identity to positions 44 to 73 set forth in SEQ ID NO:4.  
     
     
         26 . The polypeptide according to  claim 16  lacking at least one IL-1 activity.  
     
     
         27 . The polypeptide according to  claim 16  being substantially devoid of IL-1 activity.  
     
     
         28 . A CGEN-R1 protein, comprising a first portion having an amino acid sequence being at least about 90% homologous, and preferably at least about 95% homologous, to amino acids 1-68 of wild type IL-1Ra (SEQ ID NO:8), which are also amino acids 1-68 of SEQ ID NO:2; and a second portion having an amino acid sequence being at least about 80% homologous to an amino acid sequence GEWLPGKPMYVGITSLCPSVCSSMACLHKP (amino acids 69-98 of SEQ ID NO:2), wherein said first and second portions are contiguous and in sequential order.  
     
     
         29 . The CGEN-R1 protein according to  claim 28 , wherein the second portion is at least about 85% homologous to amino acids 69-98 set forth in SEQ ID NO:2.  
     
     
         30 . The CGEN-R1 protein according to  claim 28 , wherein the second portion is at least about 90% homologous to amino acids 69-98 set forth in SEQ ID NO:2.  
     
     
         31 . The CGEN-R1 protein according to  claim 28 , wherein the second portion is at least about 95% homologous to amino acids 69-98 set forth in SEQ ID NO:2.  
     
     
         32 . A CGEN-R1 protein, comprising a first portion having an amino acid sequence being at least about 90% homologous, and preferably at least about 95% homologous, to amino acids 1-43 of SEQ ID NO:4; and a second portion having an amino acid sequence being at least about 80% homologous to an amino acid sequence GEWLPGKPMYVGITSLCPSVCSSMACLHKP (amino acids 44-73 of SEQ ID NO:4), wherein said first and second portions are contiguous and in sequential order.  
     
     
         33 . The CGEN-R1 protein according to  claim 32 , wherein the second portion comprises an amino acid sequence being at least about 85% homologous to amino acids 44-73 of SEQ ID NO:4.  
     
     
         34 . The CGEN-R1 protein according to  claim 32 , wherein the second portion comprises an amino acid sequence being at least about 90% homologous to amino acids 44-73 of SEQ ID NO:4.  
     
     
         35 . The CGEN-R1 protein according to  claim 32 , wherein the second portion comprises an amino acid sequence being at least about 95% homologous to amino acids 44-73 of SEQ ID NO:4.  
     
     
         36 . A tail portion of CGEN-R1, which comprises a peptide having an amino acid sequence GEWLPGKPMYVGITSLCPSVCSSMACLHKP (amino acids 69-98 of SEQ ID NO:2), or a sequence at least about 80% homologous to said amino acid sequence.  
     
     
         37 . The tail portion of CGEN-R1 according to  claim 36 , having a sequence at least about 85% homologous to amino acids 69-98 set forth in SEQ ID NO:2.  
     
     
         38 . The tail portion of CGEN-R1 according to  claim 36 , having a sequence at least about 90% homologous to amino acids 69-98 set forth in SEQ ID NO:2.  
     
     
         39 . The tail portion of CGEN-R1 according to  claim 36 , having a sequence at least about 95% homologous to amino acids 69-98 set forth in SEQ ID NO:2.  
     
     
         40 . A bridge portion of SEQ ID NO:2, comprising a polypeptide having a length “n”, wherein n is at least about 10 amino acids in length, wherein at least two amino acids of said polypeptide comprise EG, said polyptide having a sequence starting from any of amino acid numbers 68−x to 68 of SEQ ID NO: 2; and ending at any of amino acid numbers 69+((n−2)−x) of SEQ ID NO: 2, wherein x varies from 0 to n−2; and wherein 69+((n−2)−x) cannot be larger than 98.  
     
     
         41 . The bridge portion of  claim 40 , wherein n is at least about 20 amino acids in length.  
     
     
         42 . The bridge portion of  claim 40 , wherein n is at least about 30 amino acids in length.  
     
     
         43 . The bridge portion of  claim 40 , wherein n is at least about 40 amino acids in length.  
     
     
         44 . The bridge portion of  claim 40 , wherein n is at least about 50 amino acids in length.  
     
     
         45 . The bridge portion of  claim 40 , comprising a homologous polypeptide having at least 80% homology to said polypeptide  
     
     
         46 . The bridge portion of  claim 40 , comprising a homologous polypeptide having at least 90% homology to said polypeptide.  
     
     
         47 . The bridge portion of  claim 40 , comprising a homologous polypeptide having at least 95% homology to said polypeptide.  
     
     
         48 . A bridge portion of SEQ ID NO:2, comprising a polypeptide having a length “n”, wherein n is about 4 to about 9 amino acids in length, wherein at least two amino acids of said polypeptide comprise EG, having a sequence starting from any of amino acid numbers 68−x to 68 of SEQ ID NO: 2; and ending at any of amino acid numbers 69+((n−2)−x) of SEQ ID NO: 2, wherein x varies from 0 to n−2; and wherein 69+((n−2)−x) cannot be larger than 98.  
     
     
         49 . A peptide having the amino acid sequence set forth in SEQ ID NO:27.  
     
     
         50 . An antibody which binds to at least one epitope of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, and fragments, variants and analogs thereof.  
     
     
         51 . An antibody which binds to at least one epitope of a polypeptide having an amino acid sequence according to  claim 16 .  
     
     
         52 . An antibody which binds to at least one epitope of a peptide having the amino acid sequence set forth in SEQ ID NO:27.  
     
     
         53 . An antibody-antigen complex formed from an antibody and epitope according to  claim 50 .  
     
     
         54 . An expression vector comprising the polynucleotide sequence according to  claim 1 .  
     
     
         55 . A host cell comprising the vector according to  claim 54 .  
     
     
         56 . A process for producing a polypeptide comprising; 
 a. culturing the host cell of  claim 55  under conditions suitable to produce the polypeptide encoded by said polynucleotide and;    b. recovering said polypeptide.    
     
     
         57 . A method for detecting a polynucleotide which encodes IL-Ra in a biological sample comprising the steps of: 
 a. hybridizing the polynucleotide sequence according to  claim 13  to a nucleic acid material of a biological sample, thereby forming a hybridization complex; and    b. detecting the hybridization complex, wherein the presence of the complex correlates with the presence of a polynucleotide encoding IL-1Ra in the biological sample.    
     
     
         58 . A pharmaceutical composition comprising as an active ingredient a polynucleotide sequence according to  claim 1 , and further comprising a pharmaceutically acceptable diluent or carrier.  
     
     
         59 . A pharmaceutical composition comprising as an active ingredient an expression vector according to  claim 54 , and further comprising a pharmaceutically acceptable diluent or carrier.  
     
     
         60 . A pharmaceutical composition comprising as an active ingredient a host cell according to  claim 55 , and further comprising a pharmaceutically acceptable diluent or carrier.  
     
     
         61 . A pharmaceutical composition comprising as an active ingredient a polypeptide according to  claim 16 , and further comprising a pharmaceutically acceptable diluent or carrier.  
     
     
         62 . A method for preventing, treating or ameliorating an IL-1 related disease or disorder, comprising administering to a subject in need thereof a pharmaceutical composition comprising as an an active ingredient a polypeptide according to  claim 16 .  
     
     
         63 . A method for preventing, treating or ameliorating an IL-1 related diseases or disorder, comprising administering to a subject in need thereof a pharmaceutical composition according to  claim 58 .  
     
     
         64 . A method according to  claim 62  wherein the IL-1 related disease or disorder is selected from the group consisting of acute pancreatitis; ALS; Alzheimer's disease; cachexia/anorexia; asthma; atherosclerosis; chronic fatigue syndrome; diabetes; glomerulonephritis; graft versus host rejection; hemorrhagic shock; hyperalgesia; inflammatory bowel disease; inflammatory conditions of a joint: osteoarthritis, psoriatic arthritis, rheumatoid arthritis; ischemic injury; cerebral ischemia; neurodegeneration; lung diseases; multiple myeloma; multiple sclerosis; myelogenous; leukemia; myopathies: muscle protein metabolism; osteoporosis; Parkinson's disease; chronic pain; pre-term labor; psoriasis; reperfusion injury; septic shock; side effects from radiation therapy; temporal mandibular joint disease; tumor metastasis; 
 inflammatory condition resulting from strain, sprain, cartilage damage, trauma, orthopedic surgery, and infection.    
     
     
         65 . An oligonucleotide sequence of at least 17 bases specifically hybridizable with a polynucleotide sequence according to  claim 1 , so as to direct exponential amplification of a portion thereof in a nucleic acid amplification reaction.  
     
     
         66 . An oligonucleotide having a nucleotide sequence selected from the group consisting of SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21 and SEQ ID NO:22.  
     
     
         67 . An isolated DNA sequence produced by using a pair of primers comprising a nucleotide according to  claim 65  in a DNA amplification method, wherein the primers are combined in the assay with the polynucleotide sequence as a template.